1 Department of Anesthesia and Intensive Care Medicine, Centre Hospitalier Universitaire de Liège, Liège, Belgium.
2 Center for Oxygen Research and Development, Institute of Chemistry B6a, University of Liège, Liège, Belgium.
Int J Immunopathol Pharmacol. 2017 Dec;30(4):362-370. doi: 10.1177/0394632017739530. Epub 2017 Oct 31.
Volatile anesthetics have been shown to modulate polymorphonuclear neutrophil (PMN) functions. The aim of this study was to examine the impact of clinically relevant concentrations of sevoflurane (SEVO), a volatile anesthetic, on the release of reactive oxygen species (ROS), myeloperoxidase (MPO), and elastase (EL) from human activated PMNs. For this purpose, samples of whole blood were collected from healthy volunteers and exposed in vitro to 2.3% or 4.6% SEVO in air. To assess for a stimulus-dependent effect of the volatile anesthetic, PMNs were activated using different validated protocols. Artificial stimulation of neutrophils involved either a combination of cytochalasin B (CB) and N-formyl-methionyl-leucyl-phenylalanine (fMLP) or phorbol 12-myristate 13-acetate (PMA). In addition, a combination of lipopolysaccharide (LPS) and tumor necrosis factor alpha (TNF-α) was also tested as a natural activation mean of PMNs. The production of ROS by PMNs was assessed by L-012 chemiluminescence. Total MPO and EL released in supernatant were measured by enzyme-linked immunosorbent assay (ELISA). Furthermore, degranulation of the active fraction of MPO was also measured by specific immunological extraction followed by enzymatic detection (SIEFED). Overall, SEVO enhanced the release of ROS, MPO, and EL following artificial stimulation of PMNs but the volatile anesthetic inhibited the degranulation of active MPO and EL after neutrophil exposure to LPS and TNF-α. This study highlighted that the effect of SEVO on activated PMNs is dependent on the conditions of cell stimulation. These properties should be taken into consideration in future studies investigating immunomodulatory effects of volatile anesthetics.
挥发性麻醉剂已被证明可调节多形核粒细胞 (PMN) 的功能。本研究旨在研究临床相关浓度的七氟醚 (SEVO) 对人激活的 PMN 释放活性氧 (ROS)、髓过氧化物酶 (MPO) 和弹性蛋白酶 (EL) 的影响。为此,从健康志愿者采集全血样本,并在体外暴露于空气中的 2.3% 或 4.6% SEVO。为了评估挥发性麻醉剂的刺激依赖性效应,使用不同的验证方案激活 PMN。人工刺激中性粒细胞涉及细胞松弛素 B (CB) 和 N-甲酰基-甲硫氨酸-亮氨酸-苯丙氨酸 (fMLP) 或佛波醇 12-肉豆蔻酸 13-乙酸酯 (PMA) 的组合。此外,还测试了脂多糖 (LPS) 和肿瘤坏死因子-α (TNF-α) 的组合作为 PMN 的天然激活手段。通过 L-012 化学发光法评估 PMN 产生的 ROS。通过酶联免疫吸附试验 (ELISA) 测量上清液中总 MPO 和 EL 的释放量。此外,还通过特异性免疫提取后酶检测 (SIEFED) 测量活性 MPO 脱颗粒。总体而言,SEVO 增强了 PMN 人工刺激后 ROS、MPO 和 EL 的释放,但挥发性麻醉剂抑制了 LPS 和 TNF-α 暴露后 PMN 中活性 MPO 和 EL 的脱颗粒。这项研究强调,SEVO 对激活的 PMN 的影响取决于细胞刺激的条件。在未来研究挥发性麻醉剂的免疫调节作用时,应考虑这些特性。